title: Neoplastic alterations induced in mammalian skin following mancozeb exposure using in vivo and in vitro models.
abstract: Mancozeb, ethylene(bis)dithiocarbamate fungicides, has been well documented in the literature as a multipotent carcinogen, but the underlying mechanism remains unrevealed. Thus, mancozeb has been selected in this study with the objective to decipher the molecular mechanism that culminates in carcinogenesis. We employed two-dimensional gel electrophoresis and mass spectrometry to generate a comparative proteome profile of control and mancozeb (200 mg/kg body weight) exposed mouse skin. Although many differentially expressed proteins were found, among them, two significantly upregulated proteins, namely, S100A6 (Calcyclin) and S100A9 (Calgranulin-B), are known markers of keratinocyte differentiation and proliferation, which suggested their role in mancozeb-induced neoplastic alterations. Therefore, we verified these alterations in the human system by using HaCaT cells as an in vitro model for human skin keratinocyte carcinogenesis. Upregulation of these two proteins upon mancozeb (0.5 μg/mL) exposure in HaCaT cells indicated its neoplastic potential in human skin also. This potential was confirmed by increase in number of colonies in colony formation and anchorage-independent growth assays. Modulation of S100A6/S100A9 targets, elevated phosphorylation of extracellular signal regulated kinase (ERK1/2), Elk1, nuclear factor- kappa B and cell division cycle 25 C phosphatase, and cyclin D1 and cyclooxygenase-2 upregulation was seen. In addition, PD98059 (ERK1/2 inhibitor) reduced cell proliferation induced by mancozeb, confirming the involvement of ERK1/2 signaling. Conclusively, we herein present the first report asserting that the mechanism involving S100A6 and S100A9 regulated ERK1/2 signaling underlies the mancozeb-induced neoplastic potential in human skin.
authors.affiliation: Proteomics Laboratory, Indian Institute of Toxicology Research, Council of Scientific & Industrial Research (CSIR), Lucknow 226001 (U.P), India.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Tyagi
authors.forename: Jasmine
authors.initials: J
authors.lastname: George
authors.forename: Richa
authors.initials: R
authors.lastname: Singh
authors.forename: Kulpreet
authors.initials: K
authors.lastname: Bhui
authors.forename: Yogeshwer
authors.initials: Y
authors.lastname: Shukla
mesh_terms: D000818:Animals; D015153:Blotting, Western; D040502:Calgranulin B / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000187:drug effects; D005659:Fungicides, Industrial / Q000633:toxicity*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D008297:Male; D008344:Maneb / Q000633:toxicity*; D051379:Mice; D010766:Phosphorylation / Q000187:drug effects; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D012867:Skin / Q000187:drug effects* / Q000473:pathology*; D012878:Skin Neoplasms / Q000139:chemically induced / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015038:Zineb / Q000633:toxicity*
chemical_list: D040502:Calgranulin B; D018797:Cell Cycle Proteins; D005659:Fungicides, Industrial; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C489134:S100a6 protein, mouse; D008344:Maneb; C013099:mancozeb; D015038:Zineb